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副猪嗜血杆菌EZ-Tn5转座子插入突变体库的构建及减毒株的筛选
引用本文:贺云霞,徐慧,叶飞,孙慧玲,王宏俊,龚玉梅,张莉,黄秀芬,张培君.副猪嗜血杆菌EZ-Tn5转座子插入突变体库的构建及减毒株的筛选[J].安徽农业科学,2011(13):7820-7823,7831.
作者姓名:贺云霞  徐慧  叶飞  孙慧玲  王宏俊  龚玉梅  张莉  黄秀芬  张培君
作者单位:北京市农林科学院畜牧兽医研究所;首都师范大学生命科学学院;
基金项目:国家高技术研究发展计划项目(2006AA10A206); 国家自然基金项目(31001072); 北京市农林科学院青年基金项目(QNJJ201012)
摘    要:目的]获得副猪嗜血杆菌减毒株。方法]应用转座子技术构建转座子插入突变体库,卡那抗性筛选阳性菌株,PCR扩增卡那特异片段去除假阳性,小鼠感染试验检测突变株毒力,并对获得的减毒株进行生物学特性检测。结果]所获得减毒突变菌株具有与野毒株相似的增殖能力,传代后毒力稳定,遗传学特性稳定。结论]该研究结果为进一步探讨HPS毒力因子、致病机制奠定了基础。

关 键 词:副猪嗜血杆菌  转座子  突变体库  减毒株

Construction of Mutation Library in Haemophilus parasuis by Inserting Tn5 Transposon and the Screening of Attenuated Strain
HE Yun-xia et al.Construction of Mutation Library in Haemophilus parasuis by Inserting Tn5 Transposon and the Screening of Attenuated Strain[J].Journal of Anhui Agricultural Sciences,2011(13):7820-7823,7831.
Authors:HE Yun-xia
Institution:HE Yun-xia et al(Institute of Animal Husbandry and Veterinary Medicine of Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097)
Abstract:Objective]The study aimed to obtain attenuated strain of Haemophilus parasuis.Method]Tn5 transposon technology was used to construct a library of mutants.Positive mutants were screened by kanamycin resistance.False positive were identified by PCR and then removed.Mice were infected to detect the virulence of mutants.The bionomics of attenuated strains were detected,too.Result]The attenuated mutants showed similar reproductive activity to that of wild strain.The virulence of mutants was stable after 30 pa...
Keywords:Haemophilus parasuis  Transposon  Mutation library  Attenuated strains  
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